Studies on the degradation effect of corn straw by Asian corn borer larva (Ostrinia furnacalis) digestive enzymes combined with white rot fungus (Phanerochaetc chrysosporium).

The lignin, cellulose and hemicellulose contained in corn straw need to be degraded before reuse. Therefore, it is urgent to explore a new strategy that can enhance the degradation effect of lignin, cellulose and hemicellulose. Ostrinia furnacalis is one of the corn pests feeding on corn straw, which can degrade and consume corn straw by digestive enzyme secreted in the midgut. Herein, the degradation efficiency of lignin, cellulose and hemicellulose was tested by a stain of white rot fungus combined with digestive enzyme of O. furnacalis extracted from its midgut. It was proved that the selected strain of white rot fungus could degrade lignin, cellulose and hemicellulose effectively. The contents of lignin, cellulose and hemicellulose decreased with the extension of degradation time, with the lowest level reached at 35 d with 9 ml digestive enzyme solution of O. furnacalis added. Compared with the control group, digestive enzyme of O. furnacalis could improve the degradation effect of the selected white rot fungi on lignin, cellulose and hemicellulose. The effect of degradation was enhanced with the extension of degradation time and the increase in the amount of digestive enzyme added. The results provide a new strategy and a basis for strengthening the degradation effect of white rot fungi on corn straw.

Preliminary Analysis of Transcriptome Response of Dioryctria sylvestrella (Lepidoptera: Pyralidae) Larvae Infected with Beauveria bassiana under Short-Term Starvation.

The Dioryctria genus contains several destructive borer pests that are found in coniferous forests in the Northern Hemisphere. Beauveria bassiana spore powder was tested as a new method of pest control. In this study, Dioryctria sylvestrella (Lepidoptera: Pyralidae) was used as the object. A transcriptome analysis was performed on a freshly caught group, a fasting treatment control group, and a treatment group inoculated with a wild B. bassiana strain, SBM-03. Under the conditions of 72-h fasting and a low temperature of 16 ± 1 °C, (i) in the control group, 13,135 of 16,969 genes were downregulated. However, in the treatment group, 14,558 of 16,665 genes were upregulated. (ii) In the control group, the expression of most genes in the upstream and midstream of the Toll and IMD pathways was downregulated, but 13 of the 21 antimicrobial peptides were still upregulated. In the treatment group, the gene expression of almost all antimicrobial peptides was increased. Several AMPs, including cecropin, gloverin, and gallerimycin, may have a specific inhibitory effect on B. bassiana. (iii) In the treatment group, one gene in the glutathione S-transferase system and four genes in the cytochrome P450 enzyme family were upregulated, with a sharp rise in those that were upregulated significantly. In addition, most genes of the peroxidase and catalase families, but none of the superoxide dismutase family were upregulated significantly. Through innovative fasting and lower temperature control, we have a certain understanding of the specific defense mechanism by which D. sylvestrella larvae may resist B. bassiana in the pre-wintering period. This study paves the way for improving the toxicity of B. bassiana to Dioryctria spp.

The effect of spraying bacterial and fungal solutions on Korean pine Pinus koraiensis Sieb. et Zucc. cone development and seed quality when sprayed during the flowering phase.

Korean pine is an economically essential afforestation species limited by the unreasonable collection of cones, indiscriminate use of chemical pesticides and pest damage. This study aimed to determine whether spraying bacterial or fungal solutions affected insect pests, cone development, and the seed quality of Korean pine Pinus koraiensis Sieb. et Zucc. The experiment was conducted in a forest plantation in Linkou County (Heilongjiang, China) in 2019. Four fungal strains and one bacterial strain were applied during the flowering phase of Korean pine. The results after a year and a half of study indicated that a high concentration of Bacillus thuringiensis 223176 promoted cone development, increased seed weight, and reduced the proportion of damaged cones. Under this treatment, there were 15.873% damaged cones; the seed weight reached 0.829 g, and there were 82.738% fully developed cones. Trees treated with the second most effective strain, Beauveria bassiana 122077, had 30.556% damaged cones and an average seed weight of 0.810 g. Leucanicillium antillanum 01 performed the worst in this study. The seed weight was only 0.775 g, and the damaged and fully developed cones were 52.444 and 41.773%, respectively. In summary, spraying bacterial or fungal solutions during the flowering stage of Korean pine positively impacted seed quality and effectively decreased damage by the lepidopteran species that feed on the cones and seeds in this study.

Genes involved in the Type I pheromone biosynthesis pathway and chemoreception from the sex pheromone gland transcriptome of Dioryctria abietella.

Dioryctria abietella is a coniferous seed orchard pest that can damage a series of host plants and cause huge losses to the forest economy. Sex pheromones play an important role in lepidopteran sex communication for reproduction and can be used as biological control agents to monitor and trap pests. However, the genes involved in the biosynthesis, transportation, and degradation of D. abietella sex pheromones have not been studied extensively. Transcriptome analysis of female D. abietella sex pheromone glands (PGs) revealed that 210 candidate genes might be involved in sex pheromone biosynthesis (139 genes) and chemoreception systems (71 genes). The gene expression patterns exhibited four desaturase genes (DabiDES4-7) and one fatty acid reductase gene (DabiFAR6), which were more highly expressed in sex pheromone glands than in other tissues, suggesting that these enzymes play an important role in D. abietella sex pheromone synthesis. In addition, most DabiOBPs showed high expression in antennae, but only DabiOBP4 exhibited specific expression in sex pheromone glands, suggesting that they may play many physiological roles in D. abietella. We put forth a reasonable hypothesis about type I pheromone biosynthesis pathways based on these genes identified in the D. abietella sex pheromone gland transcriptome. Our findings lay a foundation for population monitoring, mating disruption, mass trapping, and the development of ecologically acceptable management strategies.

Identification and analysis of olfactory genes in Dioryctria abietella based on the antennal transcriptome.

The coneworm Dioryctria abietella (Lepidoptera: Pyralidae) is an economy devastating pest that infests many valuable conifer species in the Holarctic regions, such as Pinus koraiensis Siebold and Zucc. The chemosensory system plays a crucial role in the mating, foraging, and ovipositing of this pest, and therefore it is desirable to identify chemosensory molecules for pest control. However, little is known at molecular level about the olfactory mechanisms in D. abietella. In the present study, we first established antennal transcriptomes of D. abietella and identified 132 putative chemosensory genes, including 15 odorant-binding proteins, 18 chemosensory proteins, 65 odorant receptors, 5 sensory neuron membrane proteins, 24 ionotropic receptors, and 5 gustatory receptors. In addition, phylogenetic trees were constructed for chemosensory genes to investigate the orthologs between D. abietella and other species of insects. Furthermore, we also compared the patterns of motifs between OBPs and CSPs using MEME. Additionally, we observed that most of DabiOBPs and DabiCSPs had the antenna-biased expression by quantitative real-time PCR (RT-qPCR), and there was a higher expression of DabiPBP1 and DabiPBP2 in male antennae than in female antennae. The binding sites of DabiPBPs (DabiPBP1, DabiPBP2) and DabiPRs (DabiOR19, DabiOR31) to the sex pheromone were predicted well by three-dimensional docking structure modelling and molecular docking. Our finding supplied a foundation for further research on the binding process of OBPs or CSPs and sensing process of ORs, SNMPs, IRs or GRs in D. abietella.

Screening of high-virulent entomopathogenic fungal strains to infect Xylotrechus rusticus larvae.

The gray tiger longicorn beetle, Xylotrechus rusticus Linnaeus (Coleoptera: Cerambycidae) is a stem-boring pest that can inhibit not only the transportation of nutrients in the trunk but also the tree growth, increasing the risk of tree breakage and causing economic losses. It is distributed in China, Iran, Turkey, Russia, Korea, Japan, and Southern Europe. This study aimed to investigate selected Beauveria strains that could be used as entomopathogenic fungi for the biological control of this pest. The high-virulence strains were screened among the selected strains by cumulative mortality, correct mortality, and lethal time to 50% mortality (LT50). These screened high-virulence strains were Bb01, CFCC83486, and CFCC81428. Bb01 exhibited 96.96% cumulative mortality, with an estimated LT50 of 3.28 days. CFCC83486 and CFCC81428 caused 89.29% and 75.74% cumulative mortality, with an estimated LT50 of 3.45 and 4.28 days, respectively. Pathogenicity at different concentrations and lethal concentration of 50% (LC50) of these high-virulence strains were investigated. The pathogenicity was found to be positively correlated with suspension concentration, and LC50 was negatively correlated with infection time. These suspensions of high-virulence strains at different concentrations were also investigated in the forest by brushing the suspensions on the poplar tree trunk infested with X. rusticus L. larvae. The most effective strain was found to be Bb01, whose cumulative mortality reached 76.33% at 1.32 × 108 conidia mL-1, followed by the strain CFCC83486, whose cumulative mortality reached 65.17% at 1.32 × 108 conidia mL-1. This study provides an important basis for using B. bassiana in the biological control of X. rusticus L.

Transcriptome analysis of abscisic acid induced 20E regulation in suspension Ajuga lobata cells.

Ajuga lobata D. Don is a medicinal plant rich in 20-hydroxyecdysone (20E), alkaloids, and other active substances. In this study, the cell suspension was incubated for 7 days, followed by the analysis on the effects of abscisic acid (ABA) on the regulation of 20E synthesis. Then A. lobata suspension cells treated with 0.15 mg/l ABA were used as material, with the Illumina technology applied for transcriptome sequencing. Digital analysis on the gene expression profile was carried out on ABA treated and control samples, respectively. Finally, transcriptomics was applied to assess the molecular response of A. lobata induced by ABA through applying transcriptomics by evaluating differentially expressed genes. The results suggested that ABA promoted 20E accumulation, while longer processing time caused cell browning. A total of 154 genes were significantly regulated after ABA treatment, with 99 up-regulated and 55 down-regulated, respectively. In addition to 20E-related pathways, the genes belonged to the ko00900 (terpenoid backbone biosynthesis) pathway (six differentially expressed genes [DEGs]), ko00100 (steroid biosynthesis) pathway (four DEGs), and ko00140 (steroid hormone biosynthesis) pathway (six DEGs). Providing a better understanding of the 20E biosynthetic pathway and its regulation, in particular in plants, this study is necessary.

ß-Ecdysterone accumulation and regulation in Ajuga multiflora Bunge suspension culture.

Ajuga multiflora Bunge cells contain ß-ecdysterone (ß-EC) that regulates the molting process of insect larvae. In this study, different conditions of culture have been studied to optimize the production of ß-EC. A. multiflora Bunge growth fitted the curve of logistic equation with one growth cycle of 17 days. The electric conductivity of medium had a negative correlation with not only the weight of dry cell but also the ß-EC accumulation, and thus, could be used for monitoring the peak of both cell growth and ß-EC accumulation. The pH value of the culture medium varied from 4.67 to 5.84 and reached the maximum at the end of the culture (on the 17th day). The relation of cell growth and nutrient consumption in A. multiflora Bunge cell suspension culture was distinctly correlated. Continuous subculture caused a reduction in ß-EC synthesis; passages 7-15, the ß-EC content declined (p < 0.05). At passage 11, the ß-EC content was only 42.72% of that at passage 5. Additives such as mevalonic acid (MVA), l-phenylalanine (l-Phe), α-pinene, terpineol, and nitric oxide (NO) in the suspension culture medium, could significantly promote the cell growth and stimulate ß-EC accumulation. The optimal concentrations of l-Phe, MVA, terpineol, and α-pinene were 0.2 mmol/l, 10 mg/l, 1 mmol/l and 6 mmol/l, respectively, with the ß-EC concentrations as 1.914 ± 0.1948 mg/g (p < 0.01), 6.012 ± 0.4252 mg/g (p < 0.01), 5.147 ± 0.4819 mg/g (p < 0.01), 2.801 ± 0.1253 mg/g (p < 0.01), respectively. The optimal concentration of sodium nitroprusside, the provider of NO, was 3 mmol/l with the ß-EC concentration 2.87 ± 0.2493 mg/g (p < 0.01). The results offer a strategy for massive production of ß-EC.

20-hydroxyecdysone accumulation and regulation in Ajuga lobata D. Don suspension culture.

Suspension culture of Ajuga lobata D. Don cells provides a method of synthesis of the phytoecdysteroid 20-hydroxyecdysone (20E) which can regulate the molting process of larvae. We characterized the culture conditions to optimize 20E production. Growth of A. lobata D. Don cells fits the logistic equation curve with a growth cycle of 19 days. Medium conductivity was negatively correlated with dry cell weight and 20E accumulation, thus could be used to determine the optimal time for cell harvest. Continuous subculture reduced 20E synthesis, but supplementing medium with 20E precursors mevalonic (MVA), α-Pinene, and nitric oxide (NO) can significantly promote cell growth and influence 20E accumulation. Combination of α-Pinene, MVA, and SNP significantly elevated 20E accumulation, thus may synergistically enhance 20E synthesis in A. lobata D. Don. The optimal concentrations of α-Pinene, MVA, and NO donor SNP in suspension culture were 50 µL L(-1), 10 mg L(-1), and 80 µmol L(-1).

Novel photodegradable insecticide W/TiO(2)/Avermectin nanocomposites obtained by polyelectrolytes assembly.

Stable and even microcrystals of Avermectin (AVM) were produced by recrystallization in presence of a stabilizer. Sequential layer growth was achieved by the layer-by-layer (LbL) self-assembly of biocompatible polyelectrolytes (PEs). The coated colloids were characterized using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). The in vitro release of Avermectin from microcapsules was studied under the simulated insect midgut conditions. W-doped TiO(2) photocatalysts were synthesized by a simple hydrothermal method, and characterized by Brunauer-Emmett-Teller (BET) surface area measurements and SEM. The photocatalytic activities of photocatalysts, which were undoped with TiO(2) and W-doped TiO(2), were evaluated by the photocatalytic oxidation degradation of AVM microcapsules in aqueous solution under UV illumination. The toxicity of the photodegradable insecticide was evaluated by the adult stage Martianus dermestoides. The results showed that AVM microcrystals which were obtained by association had a mean length of 13.8µm and a zeta potential of -34.7mV. The drug loading and encapsulation efficiency were 65.57±0.96% and 46.15±0.96%, respectively. The in vitro release experiments revealed that the polyelectrolytes prolonged the release time of the encapsulated AVM microcrystals. The sample which was prepared at 120°C with 4.0mol% W-doped amount had the highest photocatalytic activity. Toxicity of the novel photodegradable insecticide was higher in the adult stage compared to the 95% AVM as indicated by the lower LC(50) value.

Diversity and ecological distribution of endophytic fungi associated with medicinal plants.

A total of 973 isolates of endophytic fungi were recovered from 1,144 tissue fragments of the six medicinal plant species belonging to 4 families collected in the Beijing Botanical Garden. Of these isolates 778 sporulated and were identified into 21 taxa by morphological characteristics. Among the taxa 11 belonged to Coelomycetes, 6 to Ascomycetes, and 4 to Hyphomycetes. Various numbers of endophytic fungi (5-8 taxa) were obtained from each plant. Alternaria alternata was the dominant species in the 6 plants, and Microsphaeropsis conielloides was also dominant in Eucommia ulmoides. There were high colonization rates (47.9%-63.1%) and isolation rates (0.7-0.93) of endophytic fungi, and they were conspicuously higher in twigs than those in leaves in the 6 plants examined. The colonization and isolation rates of endophytic fungi increased with the twig age. The results based on the analyses of cluster and Sorenson's similarity coefficients indicated that some endophytic fungi showed a certain degree of host and tissue preference.

[Consanguinity of sixteen olethreutid moth species in northeast China based on RAPD-PCR analysis].

With RAPD-PCR technique, the phylogenetic tree of 16 olethreutid moth species in northeast China was constructed, and their consanguinity was analyzed. The results showed that the clustering of 3 Pandemis species,2 Argyrotaenia species and 4 Archips species on this phylogenetic tree was coincident with their morphological taxology, but that of some other species was not. For example, Clepsis rurinana and C. pallidana were not on the same clade, but on the clade with Pandemis cinnamomeana and Ptycholoma imitartor, respectively. Genetic distance could be an index for genus, and 0.7 was the boundary. The species genetic distance from the same genus was less than 0.7, while that from different genus was more than 0.7.

[Research advance in application of RAPD techniques in entomology].

This paper summarized the application of RAPD techniques in insect taxology and ecology, relative of species, systemic development, identification of pest, diagnosing for pest resistance, construction of molecular linkage map, and molecular assistant breeding. The problems existing in the applications of RAPD in entomology were indicated, and their countermeasures were given. It was mentioned that with the development of theories and experimental techniques, insect molecular biology would make new progress and conquer all those problems.

[Advances of studies on the effects of plant volatiles on insect behavior].

Plant volatiles show different effects on insect behaviors, which included luring, repelling and killing. After damaged by phytophagous insects, some host plants could send volatiles luring natural enemies as an indirect defense. A summarize on the new advances of plant volatiles was given. The role of plant volatiles in integrated pest management in future was also discussed.